WO1998016202B1 - Fusogenic liposome composition and method - Google Patents

Fusogenic liposome composition and method

Info

Publication number
WO1998016202B1
WO1998016202B1 PCT/US1997/018838 US9718838W WO9816202B1 WO 1998016202 B1 WO1998016202 B1 WO 1998016202B1 US 9718838 W US9718838 W US 9718838W WO 9816202 B1 WO9816202 B1 WO 9816202B1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
hydrophilic polymer
liposomes
chains
liposome
Prior art date
Application number
PCT/US1997/018838
Other languages
French (fr)
Other versions
WO1998016202A3 (en
WO1998016202A2 (en
Filing date
Publication date
Application filed filed Critical
Priority to DK97912775T priority Critical patent/DK0932391T3/en
Priority to EP97912775A priority patent/EP0932391B1/en
Priority to AT97912775T priority patent/ATE232086T1/en
Priority to BR9712230-0A priority patent/BR9712230A/en
Priority to IL12929297A priority patent/IL129292A0/en
Priority to AU49878/97A priority patent/AU715063B2/en
Priority to JP51862998A priority patent/JP2001504093A/en
Priority to CA002267904A priority patent/CA2267904C/en
Priority to DE69718924T priority patent/DE69718924T2/en
Publication of WO1998016202A2 publication Critical patent/WO1998016202A2/en
Publication of WO1998016202A3 publication Critical patent/WO1998016202A3/en
Publication of WO1998016202B1 publication Critical patent/WO1998016202B1/en

Links

Abstract

A fusogenic liposome composition for delivering a liposome-entrapped compound into the cytoplasm of a target cell is described. The liposomes have an outer surface coating of chemically releasable hydrophilic polymer chains which shield hydrophobic polymers on the liposomes outer surface. Release of the hydrophilic polymer chains exposes the hydrophobic polymers for interaction with outer cell membranes of the target cells to promote fusion of the liposome with the target cells. Also disclosed is a method for using the composition to deliver a compound to target cells, and a method for selecting suitable hydrophobic polymers for use in the composition.

Claims

AMENDED CLAIMS[received by the International Bureau on 14 July 1998 (14.07.98); original claims 1-5, 9, 11-12, 19-20 and 22-24 amended; remaining claims unchanged (4 pages)]
1. A liposome composition for fusion with a target membrane, comprising a suspension of liposomes designed for targeting to the target membrane, where each liposome (i) contains a therapeutic agent entrapped in the liposomes, and (ii) is composed of vesicle-forming lipids, a portion of the lipids derivatized by a diblock copolymer composed of a hydrophobic polymer chain covalently bound to the lipid and a hydrophilic polymer chain, the hydrophobic and hydrophilic chains being joined by a chemically releasable bond for release of the hydrophilic polymer chains and exposure of the hydrophobic polymer chains.
2. The composition of claim 1, wherein the liposomes are further composed of vesicle-forming lipids having a hydrophilic polymer chain linked to a vesicle-forming lipid via a chemically releasable bond.
3. The composition of claim 1, wherein said releasable bond is a disulfide bond or a pH sensitive chemical linkage.
4. The composition of claim 1, wherein said hydrophilic polymer chains are composed of a hydrophilic polymer selected from the group consisting of polyvinylpyrrolidone, polyvinylmethylether, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyloxazoline, polyhydroxypropylmethacrylamide, polymethacrylamide, polydimethylacrylamide, polyhydr- oxypropylmethacrylate, polyhydroxyethylacrylate, hydroxymethylcellulose, hy- droxyethylcellulose, polyethyleneglycol, and polyaspaitamide.
5. The composition of claim 4, wherein said hydrophilic polymer chains are composed of polyethylene glycol chains having a molecular weight of between 500-10,000 daltons.
6. The composition of claim 1, wherein said hydrophobic polymer is selected from the group consisting of polypropylene oxide, polyethylene, polypropylene, polycarbonate, polystyrene, polysulfone, polyphenylene oxide and polytetramethylene ether.
7. The composition of claim 6, wherein said hydrophobic polymer is polypropylene oxide having a molecular weight of between 500-3,000 daltons. 46
8. The composition of claim 1, wherein said hydrophobic polymer is a linear polymer effective to cause hemolysis of red blood cells when a water-soluble triblock copolymer containing the hydrophobic polymer and hydrophilic polymer chains joined to opposite ends of the hydrophobic polymer chains by disulfide bonds is incubated with such cells, and the incubate is treated with a reducing agent.
9. The composition of claim 1, wherein said liposomes further contain a ligand attached to a distal end of the hydrophilic polymer chains, said ligand effective for ligand- specifϊc binding to a receptor molecule on a target cell surface prior to chemical release of the hydrophilic polymer chains.
10. The composition of claim 9, wherein said ligand is selected from the group consisting of (i) folate, where the composition is intended for treating tumor cells having cell- surface folate receptors, (ii) pyridoxyl, where the composition is intended for treating virus- infected CD4+ lymphocytes, and (iii) sialyl-Lewisx, where the composition is intended for treating a region of inflammation.
11. The composition of claim 1, wherein the liposomes further include a ligand attached to the liposome surface, said ligand being effective to bind to a target cell surface receptor molecule after, but not before, chemical release of the hydrophilic polymer chains.
12. The composition of claim 1, wherein said liposomes further contain a cationic lipid effective to impart a positive liposome-surface charge, to enhance binding of liposomes to target cells after, but not before, chemical release of the hydrophilic polymer chains.
13. The composition of claim 1, wherein the agent entrapped in the lipid vesicles is a polynucleotide capable of expressing a selected protein, when taken up by a target cell.
14. The composition of claim 1, wherein the agent entrapped in the liposomes is an oligonucleotide or oligonucleotide analog effective for sequence-specific binding to cellular
RNA or DNA.
15. A method of delivering a compound to target cells in a subject comprising parenterally administering to the subject liposomes designed for reaching the target cells via the bloodstream, each liposome containing said compound in entrapped form and having an outer surface coating of chemically releasable hydrophilic polymer chains, and 47 contacting the liposomes at the target cells with a chemical agent effective to release said chains forming said surface coating, thereby to expose hydrophobic polymers on the liposome outer surface for interaction with outer cell membranes of the target cells and promote fusion of the liposome with the target cells.
16. The method of claim 15, wherein said hydrophilic polymer chains are releasably attached to the liposome via a reducible chemical linkage, and said contacting includes administering to the subject a reducing agent effective to release said chains.
17. The method of claim 16, wherein said chemical linkage is a disulfide linkage and said reducing agent is selected from the group consisting of cysteine, glutathione and ascorbate.
18. The method of claim 15, wherein each of said hydrophilic polymer chains is releasably attached to the liposome via a pH sensitive chemical linkage, and said contacting includes targeting the liposomes to a site having a pH effective to release said chains.
19. The method of claim 18, wherein said liposomes have sizes between 0.03-0.40 m for extravasation into a solid tumor.
20. The method of claim 15, wherein said liposomes further contain a ligand attached to a distal end of the hydrophilic polymer chains, said ligand effective for ligand- specific binding to a receptor molecule on a target cell surface before chemical release of the hydrophilic polymer coating.
21. The method of claim 20, wherein said ligand is selected from the group consisting of (i) folate, where the composition is intended for treating tumor cells having cell- surface folate receptors, (ii) pyridoxyl, where the composition is intended for treating virus- infected CD4+ lymphocytes, and (iii) sialyl-Lewisx, where the composition is intended for treating a region of inflammation.
22. The method of claim 15, wherein the liposomes further include a ligand attached to the liposome surface, said ligand being effective to bind to a target cell surface receptor molecule after, but not before, chemical release of the hydrophilic polymer coating.
23. The method of claim 15, wherein said liposomes further contain a cationic lipid effective to impart a positive liposome-surface charge, to enhance binding of liposomes to target cells after, but not before, chemical release of the hydrophilic polymer coating.
24. A method for screening a hydrophobic polymer for fusogenic activity with a target membrane, comprising adding to a suspension of target cells, atriblock copolymer composed of a segment of the hydrophobic polymer to be tested, and attached to each end of the polymer segment through a chemically releasable bond, a hydrophilic polymer segment effective to solubilize the hydrophobic polymer segment in the suspension, releasing said hydrophilic polymers to expose said hydrophobic segments to said target cells; and analyzing said suspension for lysis of said target cells.
25. The method of claim 24, wherein said target cells are erythrocytes.
26. The method of claim 24, wherein said releasable linkage is a disulfide linkage, and said releasing includes adding a reducing agent to the suspension.
27. The method of claim 24, wherein said hydrophilic polymer is polyethylene glycol having a molecular weight between 1,000-5,000 daltons.
28. A polymer-lipid conjugate for use in promoting fusion between target membranes, comprising a first segment composed of a hydrophilic polymer; a second segment composed of a hydrophobic polymer, said second segment joined to said first segment by a chemically releasable bond; and attached to said second segment, a vesicle-forming lipid member. 49
STATEMENT UNDER ARTICLE 19
Claim 1 is amended to include the content of original claim 2; namely that the liposomes each contain a diblock copolymer composed of a hydrophobic polymer chain and a hydrophilic polymer chain. Support for this is inherent in original claim 2, and consistent with Fig. 1 and its description.
Claim 2 is amended to direct the claim to the embodiment where the liposomes further include hydrophilic polymer chains bound to a vesicle- forming lipid via a releasable chain. Basis for this is set forth in Fig. 1 and on page 10, lines 26-27.
Claim 3 is unchanged.
Claims 4 and 5 are amended for consistency with claim 1, by changing the word "coating" to "chains".
Claims 6-8 are unchanged.
Claim 9 is amended to remove the term "unshielded" to clarify that the ligand is attached to a distal end of the hydrophilic polymer chains, as set forth in Fig. 1 and its description.
Claim 10 is unchanged.
Claims 11-12 are amended to remove the term "shielded" to clarify that the ligand (claim 11) is attached to the liposome surface and that the cationic lipid (claim 12) imparts a liposome-surface charge. The word "coating" is replaced with "chains" for consistency with amended claim 1.
Claims 13-19 and 21 are unchanged.
Claims 20. 22. 23 are amended as set forth for claims 9, 11 and 12.
Claim 24 is amended to change "hemolysis" to "lysis", for consistency with the spirit of the claim in that the hydrophobic polymer is added to a suspension of target cells.
Claims 25-28 are unchanged.
PCT/US1997/018838 1996-10-11 1997-10-10 Fusogenic liposome composition and method WO1998016202A2 (en)

Priority Applications (9)

Application Number Priority Date Filing Date Title
DK97912775T DK0932391T3 (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method
EP97912775A EP0932391B1 (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method
AT97912775T ATE232086T1 (en) 1996-10-11 1997-10-10 FUSOGENIC LIPOSOME COMPOSITION AND METHODS
BR9712230-0A BR9712230A (en) 1996-10-11 1997-10-10 Fusogenic liposome method and composition.
IL12929297A IL129292A0 (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method
AU49878/97A AU715063B2 (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method
JP51862998A JP2001504093A (en) 1996-10-11 1997-10-10 Fusion liposome compositions and methods
CA002267904A CA2267904C (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method
DE69718924T DE69718924T2 (en) 1996-10-11 1997-10-10 FUSOGENIC LIPOSOME COMPOSITION AND METHOD

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US2826996P 1996-10-11 1996-10-11
US60/028,269 1996-10-11

Publications (3)

Publication Number Publication Date
WO1998016202A2 WO1998016202A2 (en) 1998-04-23
WO1998016202A3 WO1998016202A3 (en) 1998-07-16
WO1998016202B1 true WO1998016202B1 (en) 1998-09-03

Family

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Family Applications (1)

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PCT/US1997/018838 WO1998016202A2 (en) 1996-10-11 1997-10-10 Fusogenic liposome composition and method

Country Status (15)

Country Link
US (1) US5891468A (en)
EP (1) EP0932391B1 (en)
JP (1) JP2001504093A (en)
AT (1) ATE232086T1 (en)
AU (1) AU715063B2 (en)
BR (1) BR9712230A (en)
CA (1) CA2267904C (en)
DE (1) DE69718924T2 (en)
DK (1) DK0932391T3 (en)
ES (1) ES2191833T3 (en)
HK (1) HK1047550A1 (en)
IL (1) IL129292A0 (en)
PT (1) PT932391E (en)
TW (1) TW520297B (en)
WO (1) WO1998016202A2 (en)

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